Patient-Derived Organoids from cryopreserved tissue can represent clinically relevant tools for drug testing in Biliary Cancer

Abstract

Background and aims Patient-derived organoids (PDO) are three-dimensional cultures established from cancer tissues. While PDOs can replicate morphology and drug sensitivity of cancer from given patients, their clinical applicability is limited by their lack of integration in clinical trials. We aimed to define the establishment rate of PDOs from patients with cholangiocarcinoma (CCA) and establish a protocol for the generation of PDOs from cryopreserved tissue. Methods We established a biobank of PDOs from tumour tissues (N=26). A protocol for organoid establishment from cryopreserved tissue was optimised in mice, confirmed in human PDO, and validated across institutions. Results While we started from a wide enrolment of different subtypes, including Pancreatic cancer (N=7), CCA (N=11), duodenal (N=2) and Ampullary Cancer (N=6), we refined the protocol towards biliary phenotype, with reduced efficiency rate in intestinal phenotype cancers. In total, 15 PDOs were established (58%); in the CCA cohort the efficiency rate of 75% (8 out of 11). Confirmation of organoid tumour origin was achieved through pathology review within a clinically-relevant time frame [median: 1.6 months (0.43-4.83 months)]. There was no numerical enrichment of successful PDO establishment for CCAs with poor prognostic factors such as perineural invasion, vascular invasion, nodal status and resection margins (p>0.05). Data from drug screening was ready within an average of 3 months from tissue collection (range:1.4-3.4 months). PDO from cryopreserved tissue was successful in 67% of cases and maintained the morphology, secretory functions, kinetic and dynamic growth patterns, transcriptomics, and drug sensitivity compared to organoids established from fresh tissues. Conclusions Cryopreserved tissue can facilitate the centralisation of PDO establishment and thus, the integration of biliary PDOs in multicentric trials.

Abstract of "Patient-Derived Organoids from cryopreserved tissue can represent clinically relevant tools for drug testing in Biliary Cancer" by Rae C, O'Rourke CJ, Tesini G et al., https://doi.org/10.1016/j.jhepr.2026.102083, used under its Creative Commons Attribution license (CC-BY); markup removed, otherwise unmodified.